Self-Spreading of Phospholipid Bilayer in a Patterned Framework of Polymeric Bilayer

Phospholipid bilayers spontaneously spread on a hydrophilic substrate such as glass in aqueous solution due to the energetic gain of surface wetting. This process (self-spreading) was utilized to form a patterned model biological membrane containing reconstituted membrane proteins. A mechanically stable framework of a polymerized lipid bilayer was first generated by the lithographic polymerization of a diacetylene phospholipid. Then, natural lipid membranes (fluid bilayers) were introduced into the channels between polymeric bilayers by the self-spreading from a phospholipid reservoir. The spreading velocity could be fitted into a slope of -0.5 in a double logarithmic plot versus time due to the balance between the spreading force and resistive drag. The preformed polymeric bilayer accelerated the spreading by the energetic gain of covering hydrophobic edges with a lipid bilayer. At the same time, the domains of the polymeric bilayer obstructed spreading, and the spreading velocity linearly decreased with their fractional coverage. Above the critical coverage of ca. 50%, self-spreading was completely blocked (percolation threshold) and the fluid bilayer was confined in the polymer-free regions. Nonspecific adsorption of lipids onto the surface of polymeric bilayers was negligible, which enabled a heightened signal-to-background ratio in the reconstitution and observation of membrane proteins. Self-spread bilayers had a higher density of lipids than those formed by the spontaneous rupture of vesicles (vesicle fusion), presumably due to the continual supply of lipid molecules from the reservoir. These features give the self-spreading important advantages for preparing patterned model membranes with reconstituted membrane proteins.